Injection Of Proteose Peptone Research Articles

ブリ腹腔内浸出細胞からの好中球およびマクロファージの採取

In order to prepare neutrophils and macrophages, the changes in number and composition of peritoneal exudate cells (PEC) of yellowtail after injection of Pasteurella piscicida formalin killed bacteria (FKB) or proteose-peptone, respectively, were studied. The increases of PEC after injection of P. piscicida FKB was greater than that after injection of proteose-peptone. Most of the cells in the peritoneal cavity of healthy fish were lymphocytes. After injection of any of the two substances, neutrophils, eosinophils and basophils appeared at firstly, and followed by macrophages. The percentage of the lymphocytes gradually decreased. However, the appearance of macrophages in the group injected with P. piscicida FKB was earlier than in the group injected with proteose-peptone. The peak of neutrophils in PEC was observed at 6h after injection of P. piscicida FKB and 12h after injection of proteose-peptone. The peak of macrophages in PEC was observed at 48h after injection of P. piscicida FKB and 192h after injection of proteose-peptone.According to the data, P. piscicida FKB shows in elicite the exudation of macrophages into peritoneal cavity than proteose-peptone.

Divergent changes in antimicrobial activity after immunologic activation of mouse peritoneal macrophages.

To find out whether activated macrophages display a nonspecific enhancement of antibacterial activity, we determined the intracellular killing of bacteria by peritoneal macrophages from CBA and C57BL/10 mice infected with BCG and challenged with mycobacterial antigens (purified protein derivative (PPD]. After in vivo phagocytosis, the rate of in vitro intracellular killing of Listeria monocytogenes by bacillus Calmette-Guérin (BCG)-PPD-activated macrophages from CBA mice increased by a factor of 1.7 and that of those from C57BL/10 mice by a factor of 2.0, relative to the rate in normal resident macrophages. The increased listericidal activity of BCG-PPD-activated macrophages could not have been due to an increased number of peroxidase-positive macrophages because exudate macrophages obtained after i.p. injection of proteose peptone into BCG-infected mice or PPD into control mice, killed ingested Listeria about as efficiently as normal resident macrophages did. In contrast, BCG-PPD-activated macrophages from both mouse strains killed Salmonella somewhat less efficiently and Escherichia coli and Staphylococcus aureus with the same efficiency as normal resident macrophages did. These cells, however, inhibited the intracellular replication of Toxoplasma gondii. Activated peritoneal macrophages from listeria-infected mice showed a similar increase of the rate of intracellular killing of Listeria and absence of change in rate of intracellular killing of Salmonella. Consistent with the in vitro findings, the number of viable L. monocytogenes in the spleen and liver of BCG-infected CBA and C57BL/10 mice decreased during the first 2 days after i.v. injection, whereas Salmonella typhimurium proliferated in these organs of both mouse strains. Checking the state of activation of BCG-PPD-activated macrophages showed that these cells displayed enhanced O2-consumption and H2O2 release after stimulation with phorbol myristate acetate compared with resident macrophages. The present findings show that the antimicrobial activity of immunologically activated macrophages is not uniformly increased: for certain microorganisms (L. monocytogenes, T. gondii), this effector function is enhanced, whereas for others (S. typhimurium, S. aureus, E. coli), it is not.

Parasite accessory cell interactions in murine leishmaniasis. I. Evasion and stimulus-dependent suppression of the macrophage interleukin 1 response by Leishmania donovani.

Interleukin 1 (IL 1) is a principal mediator of the host immune response to microbial challenge. Accessory cells of the monocyte-macrophage series are a major source of this cytokine and are also chronically parasitized by protozoa of the genus Leishmania. This suggests that characterization of the macrophage IL 1 response to Leishmania would increase our understanding of the regulation of host immunity to these organisms. In the present study, the macrophage IL 1 response to Leishmania donovani was examined because infections with this organism have findings consistent with parasite-specific T cell unresponsiveness. Cytokine activity was measured either by direct stimulation or by co-stimulation of thymocytes. Conditioned media from BALB/c resident peritoneal macrophages infected with amastigotes of L. donovani contained no more IL 1 than did supernatant fluids of control cells. In contrast, supernatants from cells stimulated with lipopolysaccharide or heat-killed Listeria monocytogenes had significantly increased cytokine content. Resident cells infected with L. donovani for 4 hr before being stimulated with Listeria demonstrated a suppressed IL 1 response (approximately 40% of Listeria alone) to this secondary particulate stimulus. In contrast, the secondary response of leishmania-preinfected cells to lipopolysaccharide was not affected. To examine whether accessory cell nonresponsiveness to L. donovani (with respect to IL 1) was related to the state of macrophage activation, elicited peritoneal macrophages obtained by injection of proteose peptone were also studied. These cells responded to stimulation with lipopolysaccharide and fixed Staphylococcus aureus with increases in intracellular, membrane, and secreted cytokine activities. In contrast, L. donovani failed to induce any of these activities. This was found to be the case irrespective of whether amastigotes were alive or killed or opsonized with specific antibodies. Elicited cells preinfected with Leishmania responded normally to secondary stimulation with lipopolysaccharide, but not S. aureus (64% of Staphylococcus alone). In addition, attachment and penetration of L. donovani promastigotes and their subsequent conversion to amastigotes within macrophages failed to induce IL 1 synthesis. The findings of this study indicate that L. donovani has the ability to both evade and suppress the macrophage IL 1 response. Because this monokine provides an obligatory signal during macrophage-dependent T cell activation, evasion of signal transduction for IL 1 synthesis may be related to defects in cell-mediated immunity which occur during infections with this organism.

Differential Expression of Macrophage Effector Functions: Bactericidal Versus Tumoricidal Activities

Macrophage populations may be induced to express tumoricidal or bactericidal activities following exposure to certain stimuli. An understanding of the differences in the stimulatory mechanisms and in the characteristics of the macrophages they affect will be facilitated by comparing functional activities of various macrophage populations. The experiments described here were conducted to determine whether injection of a single stimulus necessarily drives cells to express both tumoricidal and bactericidal activities or whether selected reagents can drive cells to express one activity without expressing the other. The data show that a single population of mouse or hamster peritoneal exudate cells obtained following injection of proteose peptone is bactericidal for Listeria monocytogenes and for E. coli, but is not tumoricidal for TCMK-1, Ad2HE3, or mKS-A TU-5 target cells. In contrast, peritoneal exudate cells collected after injection of Bacillus Calmette Guerin (BCG) organisms are always highly tumoricidal, and either show no effect on Listeria monocytogenes or E. coli, or are at best bacteriostatic. Data indicate that the effector cells in these assays are macrophages, that the dissociation of tumoricidal and bactericidal activity occurs over a wide dose range, and that the tumoricidal capabilities are not artifacts of the assay system. These results suggest that a given macrophage population may preferentially express tumoricidal or bactericidal activities depending on the stimulus used.

The activation state of macrophage subpopulations from a murine fibrosarcoma.

We have separated subpopulations of macrophages from an immunogenic fibrosarcoma by the technique of unit gravity velocity sedimentation. The activation state of these subpopulations was determined by measurement of the Fc receptor avidity of adherent cells, and by their 5' nucleotidase and acid phosphatase activity. The subpopulations were compared to resident peritoneal macrophages and the peritoneal macrophage subpopulations elicited by injection of proteose peptone or C. parvum. The results show that two macrophage subpopulations exist within the tumour. The smaller, peroxidase-positive population, with a sedimentation velocity of 1-5 mm/h, is similar to proteose peptone stimulated macrophages with respect to Fc receptor expression, while the other, rapidly sedimenting population (5-9 mm/h) is partially activated. However, neither population achieved the level of activation demonstrated by rapidly sedimenting, C. parvum-activated macrophages. Analysis of the enzyme activity of rapidly adherent macrophages indicated that tumour, proteose peptone or C. parvum macrophages were all activated when compared to resident peritoneal macrophages. No significant differences were found with respect to the elevated levels of acid phosphatase in the three activated macrophage populations, but the 5' nucleotidase activity of C, parvum-elicited macrophages was significantly lower than either the proteose peptone or tumour macrophages. This again demonstrated that the tumour macrophages were less activated than C. parvum macrophages. These data show that tumour-infiltrating macrophages are a heterogeneous population composed of at least two subpopulations existing in different activation states and that within the tumour microenvironment they are not capable of differentiating to the higher activation state, demonstrated by C. parvum macrophages.